TGEX-SCBlue-Zeo - Shuttle Vector for the Mammalian Expression of scFv-Fc and VHH-Fc Fusions

Product Description

Shuttle Vector for the Mammalian Expression of scFv-Fc and VHH-Fc Fusions.

The TGEX™-SCblue-Zeo vector is designed for the mammalian expression of scFv-Fc fusions after transfer of scFv fragments from the PADL™ phagemid vector series.  TGEX™-SCblue-Zeo vector enables rapid and convenient expression of scFv fragment isolated by phage display as dimeric scFv-Fc fusion with all the advantages conferred by the Fc fragment for detection using commercially available secondary antibodies. Transfer of the scFv inserts are done seamlessly through a common double-SfiI (Fig. 1). Proper recombinant clones can be isolated using a blue/white colony screening.

Figure 1. scFv Phage Display to scFv-Fc Expression. Expression of free scFv from multiple phage clones is not an easy task in bacteria and binding is often weak because of the monovalency of scFv. Using TGEX-SCblue-Zeo scFv fragments can be easily transferred to a convenient mammalian transient expression system as scFv-Fc fusion (Valadon (2006), Yoon (2012)). With yields in the 40 to 400 mg/L range, just a few ml of culture is enough to validate your antibodies in any assay using simple secondary antibodies.

Plasmid Map

Applications

• Mammalian expression of scFv-Fc or VHH-Fc fusion molecules in combination with phage display
• Selection of stable transfectants using Zeocin™

Specifications

General Characteristics:

Plasmid Size: 5917
Promoter: CMV + TPL
Leader Peptide: pelBK
Cloning Site: Double SfiI site compatible with pADL phagemid vectors
Fusion Protein: human IgG1 Fc constant region
Selection: ampicillin (bacteria), zeocin™ (mammalian)
Replication: pMB1 high copy number

Physical Characteristics:

Concentration: 0.5 µg/µl.
Product Size: 10 µg.
Buffer: DNA Conservation Buffer (Tris/HCL 5 mM, EDTA 0.1 mM, pH 8.5, sterile).
Storage Temperature: -20°C.

Citations

US Patent Application (2022). Il1rap Antibodies. US 2022/0306750 A1